Compounds that stimulate glucose utilization and methods of use

ABSTRACT

The invention provides novel compounds of the Formula (I) that stimulate rates of glucose oxidation in myocardial cells: 
                         
wherein W, Cyc, p, Y, X, Z, R, R 1 , R 2 , R 3 , R 4 , I, m and n are as defined for Formula (I) herein. The invention also relates to pharmaceutical compositions comprising compounds capable of stimulation of glucose oxidation, methods for increasing glucose oxidation rates in myocardial cells, and methods of treatment of myocardial ischemia.

This Patent Application is a divisional of U.S. Ser. No. 11/064,713, filed Feb. 23, 2005, now U.S. Pat. No. 7,524,885, which is a continuation-in-part of U.S. Ser. No. 10/313,990, filed Dec. 5, 2002, now U.S. Pat. No. 7,074,828, issued Jul. 11, 2006, which is a continuation-in-part of U.S. Ser. No. 10/181,274, filed Apr. 9, 2003 now U.S. Pat. No. 7,084,173, issued Aug. 1, 2006, which is a United States National filing under 35 U.S.C. §371 which is based on PCT/IB02/02525, filed Apr. 1, 2002, now withdrawn, the entirety of the disclosure of each of these applications which is hereby incorporated into the present application by reference.

FIELD OF THE INVENTION

The invention relates to novel compounds that stimulate rates of glucose oxidation in myocardial cells. The invention also relates to pharmaceutical compositions comprising compounds capable of stimulating glucose oxidation, methods for increasing glucose oxidation rates in myocardial cells, and methods of treatment of myocardial ischemia.

BACKGROUND OF THE INVENTION

Myocardial ischemia is a common clinical pathology that occurs in the setting of angina pectoris, acute myocardial infarction, or during cardiac surgery. Myocardial ischemia is a major clinical problem, with its complications being the major cause of mortality and morbidity in Western society.

It has been reported that stimulating glucose oxidation both during and following ischemia can benefit the ischemic heart. Br J Pharmacol 128: 197-205, 1999, Am J Physiol 275: H1533-41, 1998. Biochimica et Biophysica Acta 1225: 191-9, 1994, Pediatric Research 34: 735-41, 1993, Journal of Biological Chemistry 270: 17513-20, 1995. Biochimica et Biophysica Acta 1301: 67-75, 1996, Am J Cardiol 80: 11A-16A, 1997, Molecular & Cellular Biochemistry 88: 175-9, 1989, Circ Res 65: 378-87, 1989, Circ Res 66: 546-53, 1990, American Journal of Physiology 259: H1079-85, 1990, American Journal of Physiology 261: H1053-9, 1991, Am J Physiol Heart Circ Physiol 280: H1762-9, 2001, J Am Coll Cardiol 36: 1378-85, 2000.

To meet the high energy demands of the contracting muscle, the heart must produce a constant and plentiful supply of the free energy carrier, adenosine triphosphate (ATP). This energy is produced by the metabolism of a variety of carbon substrates, including carbohydrates such as glucose. The metabolism of fatty acid is the other major source of energy for the heart.

Glucose metabolism in the heart consists of two important pathways, namely glycolysis and glucose oxidation.

It has been shown that during ischemia (such as that produced by angina pectoris, myocardial infarction or heart surgery) the levels of circulating fatty acids in the plasma can be dramatically elevated. Am Heart J 128: 61-7, 1994. As a result, during ischemia and reperfusion the heart is exposed to high levels of fatty acids, which results in the preferential use of fatty acids as an oxidative substrate over glucose. It further has been reported that this over-reliance on fatty acids as a major source of ATP contributes to fatty acid-induced ischemic damage. This observation has sparked numerous approaches directed at switching substrate utilization back to glucose in an attempt to protect the heart from fatty acid-induced ischemic damage. J Cardiovasc Pharmacol 31: 336-44, 1998, Am Heart J 134: 841-55, 1997, Am Physiol 273: H2170-7, 1997, Cardiovasc Drugs Ther 14: 615-23, 2000, Cardiovasc Res 39: 381-92, 1998, Am Heart J 139: S115-9, 2000, Coron Artery Dis 12: S8-11, 2001, Am J Cardiol 82: 14K-17K., 1998, Molecular & Cellular Biochemistry 172: 137-47, 1997, Circulation 95: 313-5, 1997, Gen Pharmacol 30: 639-45, 1998, Am J Cardiol 82: 42K-49K., 1998, Coron Artery Dis 12: S29-33, 2001, Coron Artery Dis 12: S3-7, 2001, J Nucl Med 38: 1515-21, 1997. Current approaches that are used to manipulate myocardial energy metabolism involve either stimulating glucose metabolism directly or indirectly (i.e., inhibiting fatty acid metabolism).

Since high fatty acid oxidation rates markedly decrease glucose oxidation, one approach to increasing glucose oxidation is to inhibit fatty acid oxidation. This has proven effective both during and following ischemia, and this pharmacological approach is starting to see clinical use. Although a number of pharmacological agents designed to inhibit fatty acid oxidation have recently been developed, the direct β-oxidation inhibitor, trimetazidine, was the first anti-anginal agent widely used that has a mechanism of action that can be attributed to an optimization of energy metabolism Circulation Research. 86: 580-8, 2000.

Trimetazidine is reported to primarily act by inhibiting fatty acid oxidation, thereby stimulating glucose oxidation in the heart.

A second clinically effective agent that is reported to switch energy metabolism from fatty acid to glucose oxidation is ranolazine. It has been reported that this agent stimulates glucose oxidation secondary to an inhibition of fatty acid oxidation Circulation 93: 135-42, 1996.

The detrimental effects of fatty acids on mechanical function during and following ischemia may also be attenuated by agents that increase glucose oxidation directly. Numerous experimental studies have reported that stimulation of glucose oxidation by using dichloroacetate (DCA) following ischemia (at the expense of fatty acids) can benefit the ischemic heart. Am Heart J 134: 841-55, 1997. Although DCA is an effective compound designed to stimulate glucose oxidation, it has a short biological half-life.

Therefore, there is need to develop novel class of compounds and to identify compounds that can stimulate glucose oxidation, have long biological life, and be effective in treatment or prevention of myocardial ischemia

SUMMARY OF THE INVENTION

In one aspect, the present invention is directed to novel compounds represented by Formula (I):

wherein

-   (a) W is C₁-C₆ alkyl, halogen, optionally substituted aryl,     optionally substituted aralkyl or optionally substituted aralkenyl; -   (b) Cyc is C₃ or C₄ cycloalkyl; -   (c) p is an integer from 0 to 4; -   (d) m is 1 or 2; -   (e) Y is O, S, or NR; -   (f) if m is 1 and if p is 0, Y is O, and n is not 0, then Z is     (cyclo)alkycarbonyl or if m is 1 and if p is 0, Y is O and n is 0,     then Z is heterocycle alkyl; -   (g) X is O, S, NR, or CR³R⁴; -   (h) R is H, alkyl, aryl, or

-   -   where i is an integer from 2 to 4;

-   (i) Z is H, alkyl, heterocycle alkyl, cycloalkyl, aryl or optionally     substituted C₁-C₆ alkylcarbonyl or

when X is NR and R is

or when X is NR, R and Z may be taken together with N to form a nitrogen-containing heterocyclic ring;

-   (j) R¹ is H, alkyl or aryl; -   (k) R² is H, alkyl, aryl or ═O; -   (l) R³ and R⁴ are, independently, H, alkyl or aryl; or when X is     CR³R⁴ then R³ and R⁴, taken together with the carbon atom, may form     a heterocyclic ring; and -   (m) n is an integer from 1 to 10; or a pharmaceutically acceptable     salt, ester or prodrug thereof.

In one alternate preferred aspect, the present invention is directed to novel compounds which are represented by Formula (IIS):

wherein (a) W is C₁-C₆ alkyl, halogen, optionally substituted aryl, optionally substituted aralkyl or optionally substituted aralkenyl; (b) Cyc is C₃ or C₄ cycloalkyl; and (c) p is an integer from 1 to 4, or a pharmaceutically acceptable salt, ester or prodrug thereof. While not wishing to be bound by any particular mechanism of action, it is believed that compounds of Formula (IIS) are esterfied with CoA in vivo.

According to one embodiment, novel compounds are provided which are represented by Formula (IIIa):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

According to an alternate embodiment, provided are novel compounds represented by Formula (IIIb):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl or aryl;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl;

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

In a further embodiment, provided are novel compounds represented by Formula (IIIc):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is O, S, or NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

Another embodiment provides compounds represented by Formula (IIId):

wherein

W is C₁-C₆ alkyl, halogen or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer for 0 to 3 when Cyc is C₄ cycloalkyl or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is O;

X is NR;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

An additional embodiment is directed to novel compounds represented by Formula (IIIe):

wherein

W is aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is 1;

Y is O, S, or NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

According to one aspect, the present invention is further directed to methods for increasing or improving glucose utilization in myocardial or other types of cells, tissue or organs of warm blooded animals, especially those which are capable of high glucose metabolism (e.g., heart and other muscles). The method comprises treating cells, tissue or organs with substituted or unsubstituted cyclopropane carboxylic acid or cyclobutane carbothioic acid represented by the Formula (IIS):

wherein W, Cyc and p are as defined in connection with Formula (IIS), or a cyclopropane carboxylic acid or cyclobutane carboxylic derivative of Formula (I) or any of Formulas (IIIa) to (IIIe).

According to an alternate aspect, the present invention is also directed to pharmaceutical compositions comprising a compound according to Formula (IIS), Formula (I) or any of Formulas (IIIa) to (IIIe) and suitable pharmaceutical carriers, excipients or fillers.

According to a further aspect, the present invention is directed to a method of treating physiological conditions or disorders that may be effectively treated by increasing of cell glucose utilization. According to one embodiment, such method comprises administering to a patient in need of such treatment an effective amount of a pharmaceutical composition comprising substituted or unsubstituted cyclopropane carboxylic acid or cyclobutane carbothioic acid according to Formula (IIS) or a cyclopropane carboxylic acid or cyclobutane carboxylic acid derivative of Formula (I) and any of Formulas (IIIa) to (IIIe).

The present invention is further directed to kits including a pharmaceutical composition according to the present invention.

The methods of the present invention are applicable for treating warm blooded animal subjects, such as mammals, including humans, primates, etc.

Additional embodiments will be apparent from the Detailed Description and from the claims.

DEFINITIONS

In accordance with the present invention and as used herein, the following terms are defined to have following meanings, unless explicitly stated otherwise:

The term “alkenyl” refers to unsaturated aliphatic groups having at least one double bond.

The term “alkyl” refers to saturated aliphatic groups including straight-chain, branched-chain and cyclic (including cycloalkyl and polycyclic alkyl) groups.

The terms “alkoxy” and “alkoxyl” refer to a group having the formula, R—O—, wherein R is an alkyl group.

The term “alkoxycarbonyl” refers to —C(O)OR wherein R is alkyl.

The term “aralkenyl” refers to an alkenyl group substituted with an aryl group. Preferably the alkenyl group has from 2 to about 6 carbon atoms.

The term “aralkyl” refers to an alkyl group substituted with an aryl group. Suitable aralkyl groups include benzyl, phenethyl, and the like, all of which may be optionally substituted. Preferably the alkyl group has from 1 to about 6 carbon atoms.

The term “aryl” refers to an aromatic group which has at least one ring having a conjugated pi electron system and includes a carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted.

The term “aryloxy” refers to a group having the formula, R—O—, wherein R is an aryl group.

The term “aralkoxy” refers to a group having the formula, R—O—, wherein R is an aralkyl group.

“Biaryl” refers to phenyl substituted by carbocyclic or heterocyclic aryl as defined herein, ortho, meta or para to the point of attachment of the phenyl ring.

“Brine” refers to an aqueous saturated solution of sodium chloride.

“Carbocyclic aryl” refers to aromatic groups wherein the ring atoms on the aromatic ring are carbon atoms. Carbocyclic aryl groups include monocyclic carbocyclic aryl groups and naphthyl groups, all of which may be optionally substituted. Suitable carbocyclic aryl groups include phenyl and naphthyl. Suitable substituted carbocyclic aryl groups include indene and phenyl substituted by one to two substituents such being advantageously lower alkyl, hydroxy, lower alkoxy, lower alkoxycarbonyl, halogen, trifluoromethyl, difluoromethyl, nitro, and cyano. Substituted naphthyl refers to naphthyl, more preferably 1- or 2-naphthyl, substituted by Y1, Y2 and/or Y3 as defined in connection with formula (I) hereinabove.

“Cycloalkenyl” refers to a cyclic alkenyl group. Suitable cycloalkenyl groups include, for example, cyclopentenyl and cyclohexenyl.

“Cycloalkyl” refers to a cyclic alkyl group having at least one ring and includes polycyclic groups, including fused ring cyclic alkyl groups and bridged cycloalkyl groups. Suitable cycloalkyl groups include, for example, cyclohexyl, cyclopropyl, cyclopentyl, cycloheptyl and adamantyl.

“Cyclohexylmethyl” refers to a cyclohexyl group attached to CH₂.

“Diol” refers to chemical species having at least two hydroxy (—OH) functional groups. Diols may contain more than two hydroxy groups. “Vicinal diols” are those in which two hydroxy groups are on adjacent carbons, as found for example in pinanediol.

“Fused carbocyclic” refers to a multicyclic fused carbocyclic ring having both aromatic and non-aromatic rings. Suitable fused carbocyclic rings include fluorenyl, tetralin and the like.

“Fused carbocyclic alkyl” refers to an alkyl group substituted with a fused carbocyclic ring moiety, preferably a multicyclic fused carbocyclic ring including both aromatic and non-aromatic rings. Suitable fused carbocyclic alkyl groups include fluorenylmethyl, and the like.

The term “halogen” refers to fluorine, chlorine, bromine and iodine.

“Heteroaralkenyl” refers to an alkenyl group substituted with a heteroaryl, and includes those heterocyclic systems described in “Handbook of Chemistry and Physics”, 49th edition, 1968, R. C. Weast, editor; The Chemical Rubber Co., Cleveland, Ohio. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems. Preferably the alkenyl group has from 2 to about 6 carbon atoms.

“Heteroaralkyl” refers to an alkyl group substituted with a heteroaryl, such as picolyl, and includes those heterocyclic systems described in “Handbook of Chemistry and Physics”, 49th edition, 1968, R. C. Weast, editor; The Chemical Rubber Co., Cleveland, Ohio. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems. Preferably the alkyl group has from 1 to about 6 carbon atoms.

“Heteroaryl” refers to aromatic groups having from 1 to 14 carbon atoms and the remainder of the ring atoms are heteroatoms, and includes those heterocyclic systems described in “Handbook of Chemistry and Physics”, 49th edition, 1968, R. C. Weast, editor; The Chemical Rubber Co., Cleveland, Ohio. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems. Suitable heteroatoms include oxygen, nitrogen, and S(O)i, wherein i is 0, 1 or 2, and suitable heterocyclic aryls include furanyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, and the like.

“Heterocycle” or “Heterocyclic” refers to both heteroaryl and heterocyclo groups.

“Heterocycle alkyl” refers to an alkyl group substituted with a heterocycle group.

“Heterocyclicoxy” refers to the group —OR where R is a heterocycle group.

“Heterocyclo” refers to a reduced heterocyclic ring system comprised of carbon, nitrogen, oxygen and/or sulfur atoms, and includes those heterocyclic systems described in “Handbook of Chemistry and Physics”, 49th edition, 1968, R. C. Weast, editor; The Chemical Rubber Co., Cleveland, Ohio. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems.

“Heterocycloalkyl” refers to an alkyl group substituted with a heterocyclo group, and includes those heterocyclic systems described in “Handbook of Chemistry and Physics”, 49th edition, 1968, R. C. Weast, editor; The Chemical Rubber Co., Cleveland, Ohio. See particularly Section C, Rules for Naming Organic Compounds, B. Fundamental Heterocyclic Systems. Preferably the alkyl group has from about 1 to about 6 carbon atoms.

The term “lower” referred to herein in connection with organic radicals or groups defines such radicals or groups with one and up to and including 5 carbon atoms, preferably up to and including 4 carbon atoms, and advantageously one or two carbon atoms. Such radicals or groups may be straight chain or branched chain.

“Pharmaceutically acceptable salt” includes salts of the compounds of the present invention derived from the combination of such compounds and an organic or inorganic acid. In practice the use of the salt form amounts to use of the base form. The compounds of the present invention are useful in both free base and salt form, with both forms being considered as being within the scope of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph which depicts glucose oxidation in an isolated perfused working rat heart model at the indicated concentrations of cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester (MM054) as compared to a control.

FIG. 2 is a graph which depicts glucose oxidation in an isolated perfused working rat heart model at the indicated concentrations of cyclobutanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester (MM056) as compared to a control.

FIG. 3 is a graph which depicts glucose oxidation in an isolated perfused working rat heart model at increasing concentrations of cyclopropanecarboxylic acid, 2-isopropoxy-ethyl ester (MM070) as compared to a control.

FIG. 4 is a graph which depicts glucose oxidation in an isolated perfused working rat heart model at increasing concentrations of cyclopropanecarboxylic acid (MM001) as compared to a control.

FIG. 5 is a graph which depicts glucose oxidation in an isolated perfused working heart model at the indicated concentrations of dichloroacetate (DCA), Compound 15 (MM068), Compound 41 (MM094) and Compound 45 (MM098), compared to a control.

FIG. 6 is a graph which depicts glucose oxidation in an isolated perfused rat heart model at the indicated concentrations of DCA, cyclopropanecarboxylic acid (MM001) and cyclobutanecarboxylic acid (MM013) as compared to a control.

FIG. 7 is a graph which depicts glucose oxidation in an isolated perfused rat heart model for concentrations of cyclopropanecarboxylic acid (MM001) as compared with a control.

FIG. 8 is a graph which depicts glucose oxidation in an isolated perfused rat heart model for increasing concentrations of cyclobutanecarboxylic acid (MM013) as compared to a control.

FIG. 9 is a graph which depicts Pyruvate Dehydrogenase Kinase (PDHK) activity at 1 mM of DCA or cyclopropanecarboxylic acid (MM001) as a percent of a control, using the PDHK assay described in Example C.

FIG. 10 is a graph which depicts PDHK activity at 1 mM of DCA, cyclopropanecarboxylic acid (MM001) or the CoA ester of cyclopropanecarboxylic acid (MM053) as a percent of a control, using the PDHK assay described in Example C.

FIG. 11 depicts a schematic for a mechanism of intracellular activation of cyclopropanecarboxylic acid (MM001) to give the corresponding CoA ester (MM053).

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the present invention is directed to the use of certain compounds in methods of treatment which increase glucose utilization and which ameliorate conditions which would benefit from increased glucose utilization. Included are methods for increasing glucose utilization in a cell, tissue, or organ of a warm blooded animal comprising treating said cell, tissue or organ with a glucose utilization increasing effective amount of at least one compound represented by the formulas set forth herein. Also included are methods for treatment of physiological conditions or disorders treatable by increasing glucose utilization comprising administering to a patient in need of such treatment, an effective amount to increase glucose utilization of a pharmaceutical composition comprising at least one compound as described herein. Such compounds include the compounds represented by Formulas (I), (IIS), III and (IIIa) to (IIIe).

While not wanting to be bound by any particular mechanism of action, we have observed that compounds having activity as inhibitors of Pyruvate Dehydrogenase Kinase (PDHK) demonstrate beneficial activity in stimulating glucose oxidation. Using methods that directly measure energy metabolism in a working heart model we screened compounds described herein (such as those depicted in Tables 1, 2A and 2B and MM001 and MM013) for their ability to stimulate glucose oxidation. Structures and molecular weights for MM001 and MM013 are depicted below.

Molecular Weight MM001

86.09 MM013

100.12

As a positive control we used DCA at a concentration of 3 mM. We were able to identify compounds that stimulated glucose oxidation at 100 μM. We subsequently performed concentration curves in order to determine the potency of these compounds. Data are shown in FIG. 6:

This approach was used to test the efficacy of the compounds described herein at stimulating glucose oxidation in the intact heart. MM001 is cyclopropanecarboxylic acid, which we had identified as a stimulator of glucose oxidation in the heart. MM013 is a compound that we demonstrated to stimulate glucose oxidation.

Concentration curves for MM001 and MM013 are shown in FIG. 7 and FIG. 8, respectively.

In order to determine if the activity in stimulating glucose oxidation were, indeed, associated with activity in the inhibition of PDHK, we subjected a compound (MM001) that we had identified as having glucose oxidation stimulating activity to an in vitro PDHK assay. PDHK activity was measured as described in Example C:

Our initial results demonstrated that the compound tested did not directly inhibit PDHK in vitro. However, we had evidence to prove that PDHK activity in ex vivo hearts was inhibited (FIG. 9).

It is believed that the compounds described herein act by a prodrug mechanism in inhibiting PDHK, that is, when administered in vivo the compound undergoes an intracellular modification to produce a product that inhibits PDHK. (See FIG. 11) Thus, it is believed that the compounds described herein act as prodrugs. To demonstrate this, we modified one of the compounds (MM001) in a manner in which it is believed to be modified intracellularly. This compound (MM053) is depicted below.

When tested in the in vitro PDHK assay, MM053 was active and did inhibit PDHK activity.

Molecular Weight MM053

835.61

Thus, MM053 appears to be an active form of the compound (MM001) that inhibits PDHK directly (See FIG. 10). It is believed that the compounds described herein that stimulate glucose oxidation may be similarly modified in vivo and may be prodrugs of the corresponding CoA esters.

According to one aspect, compounds of the Formula (III) which are active in stimulating glucose oxidation are provided. In particular, the compounds represented by Formula (III) are:

wherein

W is C₁-C₆ alkyl, halogen, or aryl; Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is O, S, or NR;

X is O, S, NR, or CR³R⁴;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R is H, alkyl, aryl or

where i is an integer from 2 to 4;

R¹ is H, alkyl, or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

Certain compounds of Formula (III) are represented by Formulas (IIIa) to (IIIe).

According to one aspect, the present invention provides novel compounds which are derivatives of a cyclopropane carboxylic acid or a cyclobutane carboxylic acid. These compounds exhibit glucose oxidation stimulating activity in myocardial cells and other types of cells. The compounds according to the present invention are represented by the Formula (I):

wherein

-   (a) W is C₁-C₆ alkyl, halogen, optionally substituted aryl,     optionally substituted aralkyl or optionally substituted aralkenyl; -   (b) Cyc is C₃ or C₄ cycloalkyl; -   (c) p is an integer from 0 to 4; -   (d) m is 1 or 2; -   (e) Y is O, S, or NR; -   (f) if m is 1 and if p is 0, Y is O, and n is not 0, then Z is     (cyclo)alkycarbonyl or if m is 1 and if p is 0, Y is O and n is 0,     then Z is heterocycle alkyl; -   (g) X is O, S, NR, or CR³R⁴; -   (h) R is H, alkyl, aryl, or

-   -   where i is an integer from 2 to 4;

-   (i) Z is H, alkyl, heterocycle alkyl, cycloalkyl, aryl or optionally     substituted C₁-C₆ alkylcarbonyl or

when X is NR and R is

or when X is NR, R and Z may be taken together with N to form a nitrogen-containing heterocyclic ring;

-   (j) R¹ is H, alkyl or aryl; -   (k) R² is H, alkyl, aryl or ═O; -   (l) R³ and R⁴ are, independently, H, alkyl or aryl; or when X is     CR³R then R³ and R⁴, taken together with the carbon atom, may form a     heterocyclic ring; and -   (m) n is an integer from 1 to 10; or a pharmaceutically acceptable     salt, ester or prodrug thereof.

According to an alternate aspect, the present invention provides novel compounds according to the present invention which are represented by Formula (IIS):

wherein (a) W is C₁-C₆ alkyl, halogen, optionally substituted aryl, optionally substituted aralkyl or optionally substituted aralkenyl; (b) Cyc is C₃ or C₄ cycloalkyl; and (c) p is an integer from 1 to 4; or a pharmaceutically acceptable salt, ester or prodrug thereof.

According to one embodiment, novel compounds are provided which are represented by Formula (IIIa):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

According to an alternate embodiment, provided are novel compounds represented by Formula (IIIb):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl or aryl;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl;

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

In a further embodiment, provided are novel compounds represented by Formula (IIIc):

wherein

W is C₁-C₆ alkyl, halogen, or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer from 0 to 3 when Cyc is C₄ cycloalkyl, or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is O, S, or NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

Another embodiment provides compounds represented by Formula (IIId):

wherein

W is C₁-C₆ alkyl, halogen or aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is an integer for 0 to 3 when Cyc is C₄ cycloalkyl or p is an integer from 0 to 2 when Cyc is C₃ cycloalkyl;

Y is O;

X is NR;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

An additional embodiment is directed to novel compounds represented by Formula (IIIe):

wherein

W is aryl;

Cyc is C₃ or C₄ cycloalkyl;

p is 1;

Y is O, S, or NR;

X is O, S, NR, or CR³R⁴;

R is H, alkyl, aryl, or

where i is an integer from 2 to 4;

Z is H, alkyl, cycloalkyl, aryl or (cyclo)alkylcarbonyl or

if X is NR and R is

R¹ is H, alkyl or aryl;

R² is H, alkyl, aryl or ═O;

R³ and R⁴ are, independently, H, alkyl or aryl; and

n is an integer from 1 to 10; or a pharmaceutically acceptable salt, ester or prodrug thereof.

Certain compounds according to the present invention may be conveniently prepared from the appropriate substituted or unsubstituted cyclopropane carbonyl chloride or cyclobutane carbonyl chloride according to the following reaction scheme:

wherein W, Cyc, and p are as defined in connection with Formula (I), Y is O such that R′YH is an alcohol and

R′ is

where R¹, R², X, and n are as defined in connection with Formula (I) and Z is H, alkyl (including cycloalkyl), aryl or alkycarbonyl.

Other compounds of Formula (I) and of Formulas (IIIa) to (IIIe), including those depicted in Tables 1A, 2A and 2B, may be prepared by methods similar to those described in Examples 1 to 27 and using the appropriate starting materials.

Suitable solvents include inert organic solvents such as dichloromethane and suitable base catalysts include triethylamine and pyridine.

Reaction conditions may be varied depending on the starting materials and the desired end product. Optimization of the reaction conditions would be apparent for one of ordinary skill.

The invention further provides a method for increasing the rate of glucose oxidation and improving glucose utilization in myocardial and other cells, tissue or organs of humans and animals. It has been discovered that certain substituted cyclopropanecarbothioic acid and cyclobutanecarbothioic acid derivatives and certain substituted or unsubstituted cyclopropanecarboxylic acid and cyclobutanecarboxylic acid derivatives represented by Formula (IIS), by Formula (I) and any of Formula (IIIa) to (IIIe) can increase glucose utilization in myocardial an other types of cells, tissue or organs of warm blooded animals, including humans.

Compounds of Formula (IIS) have the structure:

wherein

W is C₁-C₆ alkyl, halogen, optionally substituted aryl, optionally substituted aralkyl or optionally substituted aralkenyl; Cyc is C₃ or C₄ cycloalkyl; and p is an integer from 1 to 4; or a pharmaceutically acceptable salt, ester or prodrug thereof.

According to one embodiment, the method according to the present invention comprises treating cells, tissue or organs of an animal with at least one compound represented by Formula (I), any of Formulas (IIIa) to (IIIe) or Formula (IIS) in an amount effective to stimulate glucose utilization. The compounds of Formula (I), Formulas (IIIa) to (IIIe) or Formula (IIS) may be delivered to the cells, tissues or organs by conventional means of administrating pharmaceutical compositions such as oral administration, injection or infusion, etc., of the compounds of the Formula (I), Formulas (IIIa) to (IIIe) or (IIS) to the animal.

The invention further provides pharmaceutical compositions comprising, as its active component, at least one compound according to the Formulas (I), (IIIa) to (IIIe) or (IIS) or their pharmaceutically acceptable salts, esters or prodrugs. Pharmaceutical compositions comprising more than one compound according to the Formulas (I), (IIIa) to (IIIe) or (IIS), their various mixtures and combinations are also contemplated to be within the scope of the present invention.

Pharmaceutical compositions or formulations include compositions and formulations conventionally used in the pharmaceutical arts and may comprise carriers and excipients compatible with oral, intravenous, intramuscular, intraarterial, intracranial, and/or intracavity administration. Suitable pharmaceutical compositions and/or formulations may further compose colloidal dispersion systems, or lipid formulations (e.g., cationic or anionic lipids), micelles, microbeads, etc.

As noted, pharmaceutical compositions of the present invention may comprise pharmaceutically acceptable and physiologically acceptable carriers, diluents or excipients. Examples of suitable carriers, diluents and excipients include solvents (aqueous or non-aqueous), solutions, emulsions, dispersion media, coatings, isotonic and absorption promoting or delaying agents, compatible with pharmaceutical administration, and other commonly used carriers known in the art.

Pharmaceutical compositions may also include carriers to protect the composition against rapid degradation or elimination from the body, and, thus may comprise a controlled release formulation, including implants and microencapsulated delivery systems. For example, a time delay material such as glyceryl monostearate or glyceryl stearate alone, or in combination with a wax, may be employed.

Pharmaceutical compositions can be formulated to be compatible with a particular route of administration. For oral administration, a composition can be incorporated with excipients and used in the form of tablets, pills or capsules, e.g., gelatin capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included in oral formulations. The tablets, pills, capsules, etc., can contain any of the following ingredients, or similar compounds: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; or a flavoring or sweetening agent.

Pharmaceutical compositions for parenteral, intradermal, or subcutaneous administration can include a sterile diluent, such as water, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.

Pharmaceutical compositions for injection include sterile aqueous solutions (where water-soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). Antibacterial and antifungal agents include, for example, parabens, chlorobutanol, phenol, ascorbic acid and thimerosal. Isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride may be included in the composition. Including an agent which delays absorption, for example, aluminum monostearate and gelatin can prolong absorption of injectable compositions.

The pharmaceutical formulations can be packaged in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the pharmaceutical carrier or excipient.

The compositions can be administered by any route compatible with a desired outcome. Thus, routes of administration include oral (e.g., ingestion or inhalation), intraperitoneal, intradermal, subcutaneous, intravenous, intraarterial, intracavity, intracranial, and parenteral. The compositions can also be administered using implants and microencapsulated delivery systems.

Compositions, including pharmaceutical formulations can further include particles or a polymeric substance, such as polyesters, polyamine acids, hydrogel, polyvinyl pyrrolidone, ethylene-vinylacetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or lactide/glycolide copolymers, polylactide/glycolide copolymers, or ethylenevinylacetate copolymers. Cyclopropanecarboxylic acid, cyclopropanecarboxylic acid and derivatives and modified forms thereof can be entrapped in microcapsules, for example, by the use of hydroxymethylcellulose or gelatin-microcapsules, or poly (methylmethacrolate) microcapsules, respectively, or in a colloid drug delivery system.

In instances where cell, tissue or organ targeting is desired, a composition of the invention can of course be delivered to the target cell, organ or tissue by injection or infusion or the like. Targeting can be achieved by injection or infusion in practicing the methods of the invention. Targeting can also be achieved by using proteins that bind to a cell surface protein (e.g., receptor or matrix protein) present on the cell or population of cell types. For example, antibodies or antibody fragments (e.g., Fab region) that bind to a cell surface protein can be included in the delivery systems in order to facilitate cell, tissue or organ targeting. Viral coat proteins that bind particular cell surface proteins can be used for targeting. For example, naturally occurring or synthetic (e.g. recombinant) retroviral envelope proteins with known cell surface protein binding specificity can be employed in the liposomes in order to intracytoplasmically deliver cyclopropanecarboxylic acid, cyclopropanecarboxylic acid and derivatives and modified forms thereof into target cells, tissue or organs. Thus, delivery vehicles, including colloidal dispersion systems, can be made to have a protein coat in order to facilitate targeting or intracytoplasmic delivery of cyclopropanecarboxylic acid, cyclopropanecarboxylic acid and derivatives and modified forms thereof.

The invention further provides a method for prophylactic and therapeutic treatments of various physiological condition or disorder treatable by increasing or improving glucose utilization in cells, tissue or organs of a patient by administering to the patient in need of such treatment, effective amounts of pharmaceutical compositions comprising substituted or unsubstituted cyclopropanecarboxylic acid, cyclopropanecarboxylic acid and cyclobutanecarboxylic acid derivative compounds represented by the Formulas (I), (IIIa) to (IIIe) and (IIS).

Disorders or conditions that can be treated with a method according to the present invention include, for example, ischemic/reperfusion injury, post myocardial infarction, angina, heart failure, a cardiomyopathy, peripheral vascular disease, diabetes, and lactic acidosis, or symptoms or side effects associated with heart surgery (e.g., open heart surgery, bypass surgery, heart transplant).

The method according to the present invention includes administering a pharmaceutical compositions comprising effective amounts of substituted or unsubstituted cyclopropanecarboxylic acid, cyclopropanecarboxylic acid and cyclobutanecarboxylic acid derivative compounds represented by the Formulas (I), (IIIa) to (IIIe) and (IIS) in a single daily dose, or the total daily dosage may be administered in divided doses several times daily. Furthermore, the pharmaceutical compositions may be administered as a single dose or over a period of time.

Patients that can be treated with the method according to the present invention include all known kind of mammals, including non human primates (apes, gibbons, chimpanzees, orangutans, macaques), companion animals (dogs and cats), farm animals (horses, cows, goats, sheep, pigs), experimental animals (mouse, rat, rabbit, guinea pig), and humans.

The dosage regiment utilizing the pharmaceutical compositions according to the present invention is selected based on various factors such as type of physiological condition to be treated, age, weight, sex of the patient, severity of the conditions to be treated, the route of administration, and particular compound contained in the pharmaceutical composition. A physician or veterinarian of ordinary skill can readily determine and prescribed the effective amount of the pharmaceutical composition to prevent or to treat the specific physiological condition.

The daily dosage may be varied over wide range and can be such that the amount of an active compound selected from a substituted cyclopropanecarbothioic acid or cyclobutanecarbothioic acid, or a substituted or an unsubstituted cyclopropanecarboxylic acid or cyclobutanecarboxylic acid derivative compound represented by any of Formulas (I), (IIS) and (IIIa) to (IIIe) is sufficient to increase glucose utilization in a cell, tissue or organ of a warm blooded animal and to achieve the desired effect of alleviating or preventing fatty acid-induced ischemic damage.

The invention provides kits containing substituted or unsubstituted cyclopropanecarboxylic acid or cyclobutanecarboxylic acid, cyclopropanecarboxylic acid or cyclobutanecarboxylic acid, substituted cyclopropanecarbothioic acid or cyclobutanecarbothioic acid and derivatives and modified forms thereof represented by the Formulas (I), (IIIa) to (IIIe) and Formula (IIS), including pharmaceutical formulations, packaged into a suitable set. A kit typically includes a label or packaging insert including instructions for use, in vitro, in vivo, or ex vivo, of the components therein.

The term “packaging material” refers to a physical structure housing the components of the kit, such as cyclopropanecarboxylic acid, cyclopropanecarboxylic acid or derivatives or modified forms thereof. The packaging material can maintain the components sterilely, and can be made of material commonly used for such purposes (e.g., paper, corrugated fiber, glass, plastic, foil, ampules, etc.). The label or packaging insert can include appropriate written instructions, for example, practicing a method of the invention.

Kits of the invention therefore can additionally include instructions for using the kit components in a method of the invention. Instructions can include instructions for practicing any of the methods of the invention described herein. Thus, for example, a kit can include a cyclopropanecarboxylic acid, cyclopropanecarboxylic acid or a derivative or modified form thereof in a pharmaceutical formulation in a container, pack, or dispenser together with instructions for administration to a human subject. Instructions may additionally include indications of a satisfactory clinical endpoint or any adverse symptoms that may occur, or any additional information required by the Food and Drug Administration for use in humans.

A kit may include instructions for increasing or improving glucose utilization in vitro, ex vivo or in vivo. In other embodiments, a kit includes instructions for treating a disorder associated with deficient or inefficient glucose utilization. In one aspect, the instructions comprise instructions for treating a subject having or at risk of having ischemic/reperfusion injury, post myocardial infarction, angina, heart failure, a cardiomyopathy, peripheral vascular disease, diabetes, or lactic acidosis. In another aspect, the instructions comprise instructions for treating a subject having or at risk of having heart surgery (e.g., open heart surgery, bypass surgery, heart transplant and angioplasty).

The instructions may be on “printed matter,” e.g., on paper or cardboard within the kit, or on a label affixed to the kit or packaging material, or attached to a vial or tube containing a component of the kit. Instructions may additionally be included on a computer readable medium, such as a disk (floppy diskette or hard disk), optical CD such as CD- or DVD-ROM/RAM, magnetic tape, electronic storage media such as RAM and ROM and hybrids of these such as magnetic/optical storage media.

Kits can additionally include a buffering agent, a preservative, or a stabilizing agent. Each component of the kit can be enclosed within an individual container and all of the various containers can be within a single package.

The present invention is further illustrated in the following examples wherein all parts, percentages, and ratios are in equivalents, all temperatures are in ° C., and all pressures are atmospheric unless otherwise indicated:

EXAMPLES Example 1 Preparation of Cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester

Triethylene glycol monomethyl ether (1.1 eq, 5.26 mmol, 0.84 ml) and triethylamine (1.1 eq, 5.26 mmol, 0.73 ml) were taken in a 10 ml round bottom flask and dichloromethane (3 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (4.78 mmol, 0.5 g, 0.43 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

A yellowish-orange solid was observed after some time. Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography, and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml) and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous sodium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellow liquid. Purification was by flash chromatography and vacuum distillation (b.p.=144° C., 3.0 mm of Hg) which afforded the pure product as a colorless liquid (527.0 mg, 48%).

The compound obtained was characterized by ¹H and ¹³C NMR, IR, and mass spectroscopy:

¹H NMR (300 MHz, CDCl₃) δ 4.2 (m, 2H), 3.68 (m, 2H), 3.64 (m, 6H), 3.52 (m, 2H), 3.36 (s, 3H), 1.62 (m, 1H), 0.99 (m, 2H), 0.84 (m, 2H); IR (CHCI₃) 2876.07, 1726.62, 1199.53, 1179.49, 1107.97 cm⁻¹; ¹³C NMR (75.5 MHz, CDCI₃) δ 174.67, 71.80, 70.44, 69.06, 63.45, 58.84, 12.65, 8.35; MS (ES, MNa⁺): calculated for C₁₁H₂₀O₅Na 255.11. found 255.1.

Example 2 Preparation of Cyclobutanoylglycine (Cyclobutanecarbonyl-amino)-acetic acid)

Methyl ester glycine hydrochloride (1 eq, 2.39 mmol, 300 mg) and pyridine (2 eq, 4.78 mmol, 0.39 ml) were suspended in (5 ml) of dichloromethane followed by addition of DMAP (1.5 eq, 218.5 mg) in one portion; the reaction mixture was stirred for 30 minutes at room temperature. After 30 minutes, cyclobutanecarbonyl chloride (2 eq, 4.77 mmol, 0.54 ml) was added slowly and the reaction mixture was stirred for 4 hours at room temperature. The solvent was evaporated in vacuo and the residue extracted with ethyl acetate. The ethyl acetate layer was dried and concentrated to dryness. The crude material obtained was purified by flash chromatography to yield pure compound A (358 mg, 87%).

To a solution of A in (6 ml) THF, was added lithium hydroxide (1.1 eq, 2.3 mmol, 2.3 ml, 1M) at room temperature and the reaction mixture was stirred for 1.5 hours. The reaction mixture was then concentrated in vacuo and acidified to pH=3 with 2N HCl. The crude product was then extracted with ethyl acetate and purified by recrystallization, using an ethyl acetate/hexane mixture. The product obtained after recrystallization was further purified by flash chromatography and again recrystallization to give the title compound B as a white solid (196 mg, 59%).

The compound obtained was characterized by ¹H and ¹³C NMR, IR, and mass spectroscopy:

¹HNMR (300 MHz, CD₃OD) δ 3.87 (s, 2H), 3.14 (quintet, 1H), 1.84-2.2 (m, 6H); IR (USCOPE) 3313.01, 3098.14, 2986.18, 2940.41, 2525.15, 2469.13, 2435.25, 1738.49, 1620.96, 1566.87, 1486.61, 1346.65, 1264.23 cm⁻¹; ¹³C NMR (75.5 MHz, CD₃OD) δ 178.20, 173.12, 41.69, 40.61, 26.12, 19.01; HRMS (ES, MNa⁺) calculated for C₇H₁₁NO₃Na 180.06311. found 180.06290.

Example 3 Preparation of Cyclobutanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester

Triethylene glycol monomethyl ether (1.1 eq, 4.64 mmol, 0.74 ml) and triethylamine (1.1 eq, 4.64 mmol, 0.65 ml) were taken in a 25 ml round bottom flask and dichloromethane (3 ml) was added. This mixture was cooled to 0° C. and then cyclobutanecarbonyl chloride (4.22 mmol, 0.5 g, 0.48 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring (vigorous reaction).

A pink colored solution was observed after some time. An extra 4 ml of dichloromethane was added to maintain proper stirring (the reaction mixture became thick). Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml) and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous sodium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish-pink liquid. The liquid was purified by flash chromatography and vacuum distillation (b.p.=189° C., 3.0 mm of Hg) to yield the pure product as a colorless liquid (679.6 mg, 65.34%).

The product was characterized by ¹H and ¹³C NMR, IR and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 4.18 (m, 2H), 3.4 (m, 2H), 3.6 (m, 6H), 3.5 (m, 2H), 3.32 (s, 3H), 3.1 (quintet, 1H), 2.2 (m, 4H), 1.86 (m, 2H); IR (CDCI₃) 2946.38, 2870.68, 1730.73, 1179.35, 1109.59 cm⁻¹; ¹³C NMR (75.5 MHz, CDCI₃) δ 175.45, 71.93, 70.58, 69.21, 63.40, 59.01, 38.0, 25.24, 18.38; MS (ES, MNa⁺): calculated for C₁₂H₂₂O₅Na 269.13. found 269.1.

Examples 4, 6 to 14, 18 and 19 General Procedure for the Preparation of Certain Cyclopropanecarboxylic Acid and Cyclobutanecarboxylic Acid Derivatives

where W, Cyc, and p are as defined for Formula (I) and Y is O such that R′—YH is an alcohol, where R′ is:

where R¹, R², X, and n are defined in connection with Formula (I) and Z is H, alkyl (including cycloalkyl), aryl or alkylcarbonyl. Suitable bases include triethylamine or pyridine. Suitable solvents include dichloromethane or other inert organic solvents.

Following the procedures described in Example 1 and Example 3 and using the appropriate starting alcohol and cycloalkyl cyclopropane carboxylic acid chloride materials, (the appropriate starting alcohols were used in place of triethylene glycol monomethyl ether), the noted cyclopropanecarboxylic acid and of cyclobutanecarboxylic acid derivatives respectively were prepared (see Table I). The compounds prepared, cycloalkyl carbonylchloride and alcohol starting materials used for their preparation and their molecular weights are summarized in Table 1.

The compounds were characterized by ¹H NMR, ¹³C NMR, IR and mass spectroscopy.

Example 5 Preparation of Cyclopropanoylalanine

The procedure of Example 2 was followed except that 2.5 equivalents of pyridine was used instead of 2 equivalents, cyclopropanecarbonyl chloride was used in place of cyclobutanecarbonylchloride and methyl ester alanine hydrochloride was used in place of methyl ester glycine hydrochloride.

Purified compound B (321 mg, 87%) was characterized by ¹H and ¹³C NMR, IR, and mass spectroscopy.

¹HNMR (300 MHz, CD₃OD) δ 8.25 (br s, 1H), 4.38 (m, 1H), 3.25 (s, 1H), 1.64 (m, 1H), 1.39 (dd, 3H), 0.7-0.9 (m, 4H); IR (USCOPE) 3323.78, 3020.25, 2645.76, 1791.56, 1729.13, 1632.33, 1537.27, 1406.24, 1281.02 cm⁻¹; ¹³C NMR (75.5 MHz, CD₃ OD) δ 176.28, 176.18, 49.38, 17.77, 14.59, 7.41, 7.33; HRMS (ES, M): calculated for C₇H₁₂NO₃ 158.08117. found 158.08123.

Example 15 Preparation of Cyclopropanecarboxylic acid 2-ethoxy-ethyl ester

2-Ethoxy-ethanol (1.1 eq, 5.26 mmol, 0.47 g, 0.51 ml) and pyridine (1.1 eq, 5.26 mmol, 0.42 g, 0.43 ml) were taken in a 25 ml round bottom flask and dichloromethane (6 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (4.78 mmol, 0.5 g, 0.43 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

An orange-yellow colored solution was observed after sometime. Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish-orange liquid. Purification was by flash chromatography and vacuum distillation (b.p.=43° C., 2.8 mm of Hg) which afforded the pure product as a colorless liquid (515.8 mg, 55.4%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

1HNMR (400 MHz, CDCl₃) δ 4.22 (m, 2H), 3.62 (m, 2H), 3.53 (q, 2H), 1.64 (m, 1H), 1.2 (t, 3H), 0.98 (m, 2H), 0.84 (m, 2H); MS (ES, M+Na): calculated for C₈H₁₄O₃Na 181.19. found 181.1; IR (CH₂Cl₂) 2976.37, 2869.55, 1728.78, 1455.55, 1177.86 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 174.76, 68.39, 66.60, 63.73, 15.17, 12.91, 8.62.

Example 16 Preparation of Cyclobutanecarboxylic acid 2-ethoxy-ethyl ester

2-Ethoxy-ethanol (1.1 eq, 4.64 mmol, 0.42 g, 0.45 ml) and triethylamine (1.1 eq, 4.64 mmol, 0.47 g, 0.65 ml) were taken in a 25 ml round bottom flask and dichloromethane (6 ml) was added. This mixture was cooled to 0° C. and then cyclobutanecarbonyl chloride (4.22 mmol, 0.5 g, 0.48 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

An orange-yellow colored solution was observed after sometime. Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellow liquid. Purification was attempted by flash chromatography and vacuum distillation (b.p.=48° C., 2.8 mm of Hg) which afforded the pure product as a colorless liquid (421.3 mg, 57.7%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (500 MHz, CDCl₃) δ 4.18 (m, 2H), 3.58 (m, 2H), 3.48 (q, 2H), 3.14 (m, 1H) 2.2 (m, 4H), 1.9 (m, 2H), 1.17 (t, 3H); MS (ES, M+Na): calculated for C₉H₁₆O₃Na 195.11. found 195.1; IR (CH₂Cl₂) 2976.99, 2949.17, 1732.12, 1444.39, 1175.39 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 175.33, 68.38, 66.58, 63.51, 38.06, 25.32, 18.47, 15.16.

Example 17 Preparation of cyclopropanecarboxylic acid 2-isopropoxy-ethyl ester

2-Isopropoxy-ethanol (1.1 eq, 5.26 mmol 0.55 g, 0.61 ml) and pyridine (1.1 eq, 5.26 mmol, 0.42 g, 0.43 ml) were taken in a 25 ml round bottom flask and dichloromethane (6 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (4.78 mmol, 0.5 g, 0.43 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

An orange-yellow colored solution was observed after sometime. An extra 2 ml of dichloromethane was added to maintain proper stirring (reaction mixture becomes thick). Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish-orange liquid. Purification was by flash chromatography and vacuum distillation (b.p.=33° C., 2.9 mm of Hg) which afforded the pure product as a colorless liquid (630.2 mg, 76.40%).

Characterization of the resulting compound was done by ¹H and ¹³C NMR, IR, and mass spectroscopy:

¹HNMR (400 MHz, CDCl₃) δ 4.2 (m, 2H), 3.6 (m, 3H), 1.65 (m, 1H), 1.15 (d, 6H), 1.0 (m, 2H), 0.85 (m, 2H); IR (CH₂CI₂) 3015.93, 2972.88, 1729.05, 1454.97, 1177.85 cm⁻¹; ¹³C NMR (125 MHz, CDCI₃) δ 174.72, 71.93, 65.95, 64.0, 22.06, 12.92, 8.54; MS (ES, MNa⁺): calculated for C₉H₁₆O₃Na 195.09. found 195.0

Example 20 Preparation of Cyclobutanecarboxylic Acid, 2-(2-cyclobutanecarbonyloxy-ethoxy)-ethyl ester

Diethylene glycol (0.5 eq, 1 mmol, 0.11 g) and pyridine (2.2 eq, 2.2 mmol, 0.174 g, 0.18 ml) were taken in a 10 ml round bottom flask and dichloromethane (4 ml) was added. This mixture was cooled to 0° C. and then cyclobutanecarbonyl chloride (2.0 mmol, 0.24 g, 0.23 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

A white thick suspension was observed after sometime. Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish liquid. Purification was by flash chromatography and vacuum distillation (b.p.=113° C., 2.8 mm of Hg) which afforded the pure product as a colorless liquid (130.0 mg, 46.42%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (500 MHz, CDCl₃) δ 4.2 (m, 4H), 3.63 (m, 4H), 3.14 (m, 2H), 1.9-2.2 (m, 12H); MS (ES, M+Na): calculated for C₁₄H₂₂O₅Na 293.15. found 293.0; IR (CH₂Cl₂) 2949.36, 2869.67, 1731.42, 1445.39, 1174.72 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 175.26, 69.16, 63.32, 63.27, 38.05, 25.33, 18.49.

Example 21 Preparation of Cyclopropanecarboxylic acid, 2-[2-(2-cyclopropanecarbonyloxy-ethoxy)-ethoxy]-ethyl ester

Triethylene glycol (1.6 mmol, 0.24 g) and pyridine (2.2 eq, 3.52 mmol, 0.28 g, 0.28 ml) were taken in a 10 ml round bottom flask and dichloromethane (5 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (3.4 mmol, 0.36 g, 0.31 ml) was added in a dropwise fashion maintaining the temperature at 0° C. with constant stirring.

A white, thick suspension was observed after sometime. Stirring was continued for 1 hour at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish liquid. Purification was by flash chromatography and vacuum distillation (b.p.=127° C., 2.8 mm of Hg) which afforded the pure product as a colorless liquid (234.5 mg, 50.97%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (500 MHz, CDCl₃) δ 4.2 (m, 4H), 3.68 (m, 4H), 3.64 (br s, 4H), 1.62 (m, 2H), 0.97 (m, 4H), 0.84 (m, 4H); MS (ES, M+Na): calculated for C₁₄H₂₂O₆Na 309.14. found 309.0; IR(CH₂Cl₂) 3015.01, 2951.60, 2873.12, 1726.69, 1454.28, 1177.84 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 175.0, 70.0, 69.0, 63.0, 13.0, 8.0.

Example 22 Preparation of Cyclobutanecarboxylic acid, 2-[2-(2-cyclobutanecarbonyloxy-ethoxy)-ethoxy]-ethyl ester

The procedure described in Example 21 was followed using the following amounts of these reagents: triethylene glycol (1.6 mmol, 0.24 g), pyridine (2.2 eq, 3.52 mmol, 0.28 g, 0.28 ml); and cyclobutanecarbonyl chloride (3.4 mmol, 0.40 g, 0.39 ml).

The compound was characterized by NMR (¹H and ¹³C), IR and mass spectroscopy:

¹HNMR (500 MHz, CDCl₃) δ 4.2 (m, 4H), 3.68 (m, 4H), 3.62 (br s, 4H), 3.14 (m, 2H), 2.1-2.3 (m, 8H), 1.8-2.0 (m, 4H); MS (ES, M+Na): calculated for C₁₆H₂₆O₆Na 337.14. found 337.0; IR (CH₂Cl₂) 2948.71, 2869.27, 1731.17, 1445.80, 1175.60 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 175.29, 70.59, 69.27, 63.39, 38.06, 25.33, 18.49.

Example 23 Preparation of Cyclopropanecarboxylic acid 2-[{2-[bis(2-cyclopropanecarbonyloxy-ethyl)-amino]-ethyl}-(2-cyclopropanecarbonyloxy-ethyl)-amino]ethyl ester

A solution of the diamine tetra-ol (1 eq, 4.23 mmol, 1.0 g), pyridine (1.0 eq, 4.23 mmol, 0.33 g, 0.34 ml) and triethylamine (5.0 eq, 0.021 mol, 2.12 g, 2.90 ml) was taken in a 25 ml round bottom flask and toluene (10 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (0.019 mol, 1.98 g, 1.72 ml) was added in one shot with vigorous stirring, maintaining the temperature at 0° C.

A yellowish-white thick suspension was observed after sometime. Stirring was continued for 15 minutes at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, and extracted with ethyl acetate (2×25 ml). The ethyl acetate layer was washed with brine (1×20 ml), dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish liquid. Purification was by flash chromatography which afforded the pure product as a colorless liquid (900.0 mg, 42.0%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 4.13 (t, 8H), 3.3 (m, 3H), 2.82 (t, 8H), 2.68 (s, 4H) 1.64 (m, 4H), 0.88 (m, 16H); MS (ES, M+H): calculated for C₂₆H₄₀N₂O₈+H 509.29. found 509.0; IR (CH₂Cl₂) 3014.69, 2958.19, 2826.89, 1725.90, 1452.31, 1173.00 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 174.59, 62.62, 53.55, 53.29, 12.92, 8.50.

Example 24 Preparation of Cyclopropanecarboxylic Acid (2-isopropoxy-ethyl)-amide

A solution of the amino ether (1.1 eq, 5.26 mmol, 0.54 g, 0.64 ml), pyridine (0.5 eq, 2.39 mmol, 0.19 g, 0.19 ml), triethylamine (1.1 eq, 5.26 mmol, 0.53 g, 0.73 ml) was taken in a 10 ml round bottom flask and dichloromethane (6 ml) was added. This mixture was cooled to 0° C. and then cyclopropanecarbonyl chloride (4.78 mmol, 0.5 g, 0.43 ml) was added in a dropwise fashion, maintaining the temperature at 0° C.

A white precipitate was observed after sometime. Stirring was continued for 30 minutes at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with water (1×10 ml), brine (2×10 ml), dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a colorless liquid. Purification was by flash chromatography and vacuum distillation (b.p.=106° C., 1.4 mm of Hg) which afforded the pure product as a colorless liquid (493.8 mg, 60.22%).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 6.0 (br s, 1H), 3.57 (m, 1H), 3.44 (m, 4H), 1.32 (m, 1H) 1.14 (d, 6H), 0.94 (m, 2H), 0.7 (m, 2H); MS (EI, M⁺): calculated for C₉H₁₇NO₂ 171.12. found 171.12; IR (CH₂Cl₂) 3299.39, 3092.61, 2971.98, 2868.14, 1644.61, 1552.31, 1197.34 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 173.41, 71.76, 66.68, 39.83, 22.10, 14.72, 7.07.

Example 25 Preparation of (±)-trans-2-Phenyl-cyclopropanecarboxylic acid 2-isopropoxy-ethyl ester

2-Isopropoxy-ethanol (1.1 eq, 3.04 mmol, 0.32 g, 0.35 ml), pyridine (0.5 eq, 1.38 mmol, 0.11 g, 0.11 ml), triethylamine (1.1 eq, 3.04 mmol, 0.31 g, 0.43 ml) were taken in a 10 ml round bottom flask and dichloromethane (6 ml) was added. This mixture was cooled to 0° C. and then (±)-trans-2-phenyl-cyclopropanecarbonyl chloride (2.76 mmol, 0.5 g, 0.43 ml) was added in a dropwise fashion, maintaining the temperature at 0° C.

A white precipitate was observed after sometime. Stirring was continued for 30 minutes at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish liquid. Purification was by flash chromatography which afforded the pure product as a colorless liquid (450.0 mg, 65.0%). The above compound is racemic as determined by chiral HPLC (chiralcel OJ column) 2% Isopropanol in hexane. UV λ_(max)=278 nm (flow=1 ml/min).

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 7.07-7.27 (m, 5H), 4.22 (m, 2H), 3.61 (m, 3H), 2.51 (m, 1H) 1.93 (m, 1H), 1.58 (m, 1H), 1.26 (m, 1H), 1.53 (d, 6H); MS (EI, M⁺): calculated for C₁₅H₂₀O₃ 248.14. found 248.14; IR (CH₂Cl₂) 3063.09, 3030.28, 2971.95, 2867.20, 1727.02, 1151.71 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 173.27, 139.92, 128.36, 126.39, 126.10, 72.01, 65.94, 64.33, 26.44, 24.17, 22.10, 17.26.

Example 26 Preparation of (±)-trans-2-Phenyl-cyclopropanecarboxylic acid 2-ethoxy-ethyl ester

This compound was prepared using the procedure described in Example 25 and using the following reagents: 2-ethoxy-ethanol (1.1 eq, 3.05 mmol, 0.27 g, 0.30 ml) pyridine (0.5 eq, 1.39 mmol, 0.11 g, 0.11 ml), triethylamine (1.1 eq, 3.05 mmol, 0.31 g, 0.43 ml); and (±)-trans-2-phenyl-cyclopropanecarbonyl chloride (2.77 mmol, 0.5 g, 0.43 ml).

The compound was characterized by NMR (¹H and ¹³C), IR and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 7.06-7.28 (m, 5H), 4.24 (m, 2H), 3.62 (m, 2H), 3.52 (q, 2H) 2.51 (m, 1H), 1.94 (m, 1H), 1.58 (m, 1H), 1.29 (m, 1H), 1.19 (t, 3H); MS (EI, M⁺): calculated for C₁₄H₁₈O₃ 234.12. found 234.12; IR (CH₂Cl₂) 2975.07, 2868.63, 1726.37, 1175.66 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 173.26, 139.90, 128.37, 126.40, 126.08, 68.36, 66.65, 64.04, 26.48, 24.13, 17.35, 15.2

Example 27 Preparation of 1-Phenyl-cyclopropanecarboxylic acid 2-ethoxy-ethyl ester

1-Phenyl-cyclopropanecarboxylic acid (0.5 g, 3.1 mmol), was dissolved in thionyl chloride (10 eq, 0.031 mol, 3.68 g, 2.3 ml), and refluxed at 80° C. for 1.5 hours. Then excess of thionyl chloride was evaporated on the rotary evaporator which yielded a dark-yellowish liquid (1-phenyl-cyclopropanecarbonyl chloride A), which was then cooled to 0° C., under argon.

2-Ethoxy-ethanol (1.1 eq, 3.41 mmol, 0.31 g, 0.33 ml) and pyridine (1.2 eq, 3.41 mmol, 0.27 g, 0.28 ml) was taken in a 25 ml round bottom flask and dichloromethane (10 ml) was added. This mixture was cooled to 0° C. and then A was added in a dropwise fashion maintaining the temperature at 0° C. Stirring was continued for 30 minutes at 0° C. The reaction was monitored by thin layer chromatography and then quenched with saturated ammonium chloride solution. The mixture was then transferred to a separatory funnel, washed with 5% sodium bicarbonate (2×5 ml), 1:1 hydrochloric acid (2×5 ml), and then with brine (5 ml). The dichloromethane layer was separated from the aqueous layer, dried over anhydrous magnesium sulphate, filtered, and evaporated in vacuo to give the title product as a pale yellowish liquid. Purification was by flash chromatography and vacuum distillation (b.p.=93° C., 2.4 mm of Hg), which afforded the pure product as a colorless liquid (437.7 mg, 60.62%)

Characterization was done by NMR (¹H and ¹³C), IR, and mass spectroscopy:

¹HNMR (300 MHz, CDCl₃) δ 7.2-7.35 (m, 5H), 4.14 (m, 2H), 3.51 (m, 2H), 3.35 (q, 2H) 1.60 (dd, 2H), 1.18 (dd, 2H), 1.1 (t, 3H); MS (EI, M⁺): calculated for C₁₄H₁₈O₃ 234.12. found 234.12; IR (CH₂Cl₂) 2958.04, 2837.41, 1676.09, 1600.65, 1288.07 cm⁻¹; ¹³CNMR (125 MHz, CDCl₃) δ 174.28, 139.40, 130.41, 127.97, 127.00, 68.11, 66.57, 64.43, 29.19, 16.59, 15.23.

Note:

For the compounds of Examples 1 to 27, the solvent system used for flash chromatography was ethyl acetate/hexane, unless otherwise specified.

TABLE 1A Starting Molecular Carbonyl Starting R′- Example Compound Weight Chloride YH compound  1 MM054

232.28 P* 2-[2-(2- Methoxy- ethoxy)- ethoxy]- ethanol  2 MM055

157.17 B** Methyl ester glycine hydrochloride (Amino Acid)  3 MM056

246.31 B 2-[2-(2- Methoxy- ethoxy)- ethoxy]- ethanol  4 MM057

264.31 P 2-(2- Benzyloxy- ethoxy)- ethanol  5 MM058

157.17 P Methyl ester alanine hydrochloride (Amino Acid)  6 MM059

278.34 B 2-(2- Benzyloxy- ethoxy)- ethanol  7 MM060

244.32 B 2-(2-Butoxy- ethoxy)- ethanol  8 MM061

216.27 B 2-(2-ethoxy- ethoxy)- ethanol  9 MM062

201.26 P 2-(2- dimethylamino- ethoxy)- ethanol 10 MM063

215.29 B 2-(2- dimethylamino- ethoxy)- ethanol 11 MM064

258.35 P 2-(2- hexyloxy- ethoxy)- ethanol 12 MM065

272.39 B 2-(2- hexyloxy- ethoxy)- ethanol 13 MM066

188.23 P 2-(2-methoxy- ethoxy)- ethanol 14 MM067

202.25 B 2-(2-methoxy- ethoxy)- ethanol 15 MM068

158.20 P 2-ethoxy- ethanol 16 MM069

172.23 B 2-ethoxy- ethanol 17 MM070

172.23 P 2-Isopropoxy- ethanol 18 MM071

186.25 B 2-Isopropoxy- ethanol 19 MM072

242.27 P 2-(2-Hydroxy- ethoxy)- ethanol 20 MM073

270.32 B 2-(2-Hydroxy- ethoxy)- ethanol 21 MM074

286.32 P 2-[2-(2- Hydroxy- ethoxy)- ethoxy]- ethanol 22 MM075

314.37 B 2-[2-(2- Hydroxy- ethoxy)- ethoxyl- ethanol 23 MM076

508.60 P N,N,N′,N′ Tetrakis(2- hydroxyethyl) ethyl- enediamine 24 MM077

171.24 P 2-Amino- ethyl isopropyl ether 25 MM078

248.32 trans-2- phenyl- cyclopro- panecar- bonyl chloride 2- Isopropoxy- ethanol 26 MM079

234.29 trans-2- phenyl- cyclopro- panecar- bonyl chloride 2-ethoxy- ethanol 27 MM080

234.29 1-phenyl- cyclo- propane- carbonyl chloride 2-ethoxy- ethanol a. P* Cyclopropanecarbonyl chloride b. B** Cyclobutanecarbonyl chloride

The compounds of Table 2B may be prepared using methods within the purview of one of skill in the art, including using methods similar and/or analogous to those described in Examples 1 to 27 and in the Detailed Description of the Invention and using the appropriate starting materials.

Example A Glucose Oxidation Stimulation in Untreated Hearts and Myocardial Cells Treated with Cyclopropanecarboxylic Acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester

Rat hearts were cannulated for isolated working heart 60 minute aerobic perfusions as described in J Pharmacol Exp Ther. 1993; 264:135-144, the entire disclosure of which is incorporated herein by reference.

Male Sprague-Dawley rats (0.3-0.35 kg) were anesthetized with pentobarbital sodium (60 mg/kg IP) and hearts were quickly excised, the aorta was cannulated and a retrograde perfusion at 37° C. was initiated at a hydrostatic pressure of 60 mm Hg. Hearts were trimmed of excess tissue, and the pulmonary artery and the opening to the left atrium were then cannulated. After 15 min of Langendorff perfusion, hearts were switched to the working mode by clamping the aortic inflow line from the Langendorff reservoir and opening the left atrial inflow line. The perfusate was delivered from an oxygenator into the left atrium at a constant preload pressure of 11 mm Hg. Perfusate was ejected from spontaneously beating hearts into a compliance chamber (containing 1 ml of air) and into the aortic outflow line. The afterload was set at a hydrostatic pressure of 80 mm Hg. All working hearts were perfused with Krebs'-Henseleit solution containing calcium 2.5 mmol/L, glucose 5.5 mmol/L, 3% bovine serum albumin (fatty acid free, initial fractionation by heat shock, Sigma), and with 1.2 mmol/L palmitate. Palmitate was bound to the albumin as described in J Bio Chem. 1992; 267:3825-3831, the entire disclosure of which is incorporated herein by reference.

The perfusate was recirculated, and pH was adjusted to 7.4 by bubbling with a mixture containing 95% O₂ and 5% CO₂.

Spontaneously beating hearts were used in all perfusions. Heart rate and aortic pressure were measured with a Biopac Systems Inc. blood pressure transducer connected to the aortic outflow line. Cardiac output and aortic flow were measured with Transonic T206 ultrasonic flow probes in the preload and afterload lines, respectively. Coronary flow was calculated as the difference between cardiac output and aortic flow. Cardiac work was calculated as the product of systolic pressure and cardiac output.

Measurement of Glucose Oxidation: Glucose oxidation was measured simultaneously by perfusing hearts with [U—¹⁴C] glucose according to the procedures discussed in Saddik M, et al., J Bio Chem. 1992; 267:3825-3831. The entire disclosure of this reference is incorporated herein by reference. The total myocardial ¹⁴CO₂ production was determined at 10-min intervals from the 60-min aerobic period. Glucose oxidation rates were determined by quantitative measurement of ¹⁴CO₂ production as described in Barbour R L, et al., Biochemistry. 1984; 1923: 6503-6062. The entire disclosure of this reference is incorporated herein by reference. ¹⁴CO₂ production for the control group were compared with the ¹⁴CO₂ production for the group treated with cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester. Results are shown on FIG. 1 and TABLE 2A.

Example B (1) Glucose Oxidation Stimulation in Untreated Hearts and Myocardial Cells Treated with Cyclobutanecarboxylic Acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester

The procedure of Example A for was followed except that cyclobutanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester in 1 μM, 10 μM, 100 μM and 1000 μM amounts was added to the buffer in place of the cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester. The results are illustrated in FIG. 2 and TABLE 2A.

(2) Glucose Oxidation Stimulation in Untreated Hearts and Myocardial Cells Treated with Cyclopropanecarboxylic Acid, 2-isopropoxy ethyl ester

The procedure of Example A was followed except that cyclopropanecarboxylic acid, 2-isopropoxy-ethyl ester in 1 μM, 10 μM, 100 μM and 1000 μM amounts was added to the buffer in place of the cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester. The results are illustrated in FIG. 3 and TABLE 2A.

(3) Glucose Oxidation Stimulation in Untreated Hearts and Myocardial Cells Treated with Various Cyclopropanecarboxylic Acid and Cyclobutanecarboxylic Acid Derivatives

The procedure of Example A was followed except that various cyclobutanecarboxylic acid derivatives, cyclopropanecarboxylic acid derivatives and cyclobutanecarboxylic acid in the amounts of 100 μM or 1000 μM was added to the buffer in place of the cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester. The results are illustrated in TABLE 2A and TABLE 2B.

(4) Glucose Oxidation Stimulation in Untreated Hearts and Myocardial Cells Treated with Cyclopropanecarboxylic Acid

The procedure of Example A was followed except that cyclobutanecarboxylic acid the amounts of 0.001 μM, 0.01 μM, 01 μM, 1 μM, 10 μM, and 100 μM was added to the buffer in place of the cyclopropanecarboxylic acid, 2-[2-(2-methoxy-ethoxy)-ethoxy]-ethyl ester. The results are illustrated in FIG. 4.

TABLE 2A Glucose Compound Screening Oxida- of Concentra- tion Example tion (% above No. Compound (μM) control)  1 MM054

100 102%  2 MM055

1000 μM 58%  3 MM056

100 μM 54%  4 MM057

100 μM 104%  5 MM058

1000 μM 40%  6 MM059

100 μM 68%  7 MM060

100 μM 65%  8 MM061

Not screened  9 MM062

100 μM 77% 10 MM063

100 μM 41% 11 MM064

100 μM 83% 12 MM065

100 μM 0% 13 MM066

100 μM 20% 14 MM067

100 μM 50% 15 MM068

100 μM 416% 16 MM069

100 μM 162% 17 MM070

100 μM 208% 18 MM071

100 μM 97% 19 MM072

100 μM 97% 20 MM073

100 μM 243% 21 MM074

100 μM 228% 22 MM075

100 μM 184% 23 MM076

100 μM 274% 24 MM077

100 μM 217% 25 MM078

100 μM 200% 26 MM079

not screened 27 MM080

not screened

TABLE 2B Screening Glucose Concentra- Oxidation Molecular tion (% above No. Compound Weight (μM) control) 27 MM080

234.29 100 μM 148% 28 MM081

172.22 100 μM 121% 29 MM082

214.30 100 μM 110% 30 MM083

144.17 100 μM 197% 31 MM084

274.31 100 μM 197% 32 MM085

172.22 100 μM 150% 33 MM086

241.11 100 μM 117% 34 MM087

336.42 100 μM 112% 35 MM088

520.53 100 μM 205% 36 MM089

211.28 100 μM 202% 37 MM090

174.22 100 μM 251% 38 MM091

187.26 100 μM 176% 39 MM092

199.25 100 μM 233% 40 MM093

214.26 100 μM 260% 41 MM094

234.25 100 μM 389% 42 MM095

182.24 100 μM 314% 43 MM096

264.32 100 μM 93% 44 MM097

308.37 100 μM 125% 45 MM098

198.22 100 μM 358% 46 MM099

350.41 100 μM 93%

Example C Pyruvate Dehydrogenase Kinase (PDHK) Inhibition Assay

This assay is based on the method of Jackson, et al. Biochem J. 334:203-711 (1998). It is an adaptation of a pyruvate dehydrogenase (PDH) assay in which measures NADH (@ 340 nm) formed when pyruvate is converted to acetyl CoA. PDHK activity is measured as the amount of PDH activity remaining after ATP activation of PDHK, which in turn deactivates PDH. Pyruvate dehydrogenase enzyme complex (PDC) is purchased from Sigma. It contains intrinsic pyruvate dehydrogenase kinase (PDK) activity.

1) PDC is pre-incubated in Buffer A (40 mM MOPS, pH 7.2, 0.5 mM EDTA, 30 mM KCl. 1.5 mM MgCl₂, 0.25 mM acetyl CoA, 0.05 mM NADH, 2 mM dithiothreitol, 10 mM NaF) for 40 minutes at 37° C. This step increases total PDH activity.

2) The PDK reaction is started by adding 45.5 μl of pre-incubated enzyme (step 1) to 54.5 μl of PDK reaction solution (1.8× buffer A, 55 μM ADP±100 μM ATP,±drug). The reaction is run for 3 minutes@37° C. and then stopped by adding 10 μl of Stop solution (55 mM ADP, 55 mM pyruvate).

3) PDH activity is then assayed by adding 90 μl of Buffer B (120 mM Tris-HCl, pH 7.8, 0.61 mM EDTA, 0.73 mM MgCl2, 2.2 mM cocarboxylase, 11 mM β-mercaptoethanol, 2.2 mM NAD, 2.2 mM pyruvate, 1.1 mM coenzyme A) and read at 340 nm kinetically for 2 minutes.

4) Drug inhibition levels are compared to the control reaction (no added drug) which is considered to have 100% kinase activity.

(a) Pyruvate Dehydrogenase Kinase Assay Solutions

Buffer A (10×)

-   -   40 ml 1 M MOPS     -   1 ml of 0.5 M EDTA     -   10 ml of 3 M KCl     -   10 ml of 150 ml MgCl2     -   10 ml of 200 mM DTT

This buffer is made and kept at 4° C. On the day of the assay 7.1 ml of Buffer A (10×) is added to 1 ml of 2.5 mM acetyl CoA. 1 ml of 0.5 mM NADH, and 0.9 ml water added make Buffer A (1×)

Freshly made solutions:

-   -   2.5 mM Acetyl CoA     -   0.5 mM NADH     -   550 mM ADP     -   1 mM ATP     -   22 mM cocarboxylase     -   22 mM NAD     -   11 mM Coenzyme A         (b) PDK Reaction Solution

Solution Volume Buffer A (10X) 50 μl 550 μM ADP 27.8 μl 1 mM ATP 27.8 μl (0) Drug (or DMSO) 55.5 μl H₂O 117 μl Total Volume 277.8 μl ( ) indicates volumes for kinase negative reactions used to determine total PDH activity. (c) PDK Stop Solution

Solution Volume 550 mM ADP  150 μl 550 mM pyruvate  150 μl DD H₂O 1200 μl Total Volume 1500 μl

A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. 

1. A method for increasing glucose utilization in a cell, tissue or organ of a warm blooded animal which comprises treating said cell, tissue or organ with a glucose utilization effective amount of a compound selected from the group consisting of


2. The method of claim 1, wherein the compound is: MM090


3. The method of claim 1, wherein the compound is: MM091


4. The method of claim 1, wherein the compound is: MM095 